An intensive investigation of a soluble, cytochrome P-450-dependent alkyl-chain monooxygenase isolated from Bacillus megaterium is being undertaken. This multi-enzyme complex has been previously shown to hydroxylate fatty acids, amides and alcohols of chain-lengths C12 to C18 in the omega-1, omega-2 and omega-3 positions. The major research objectives include: 1. Purification and characterization of both the native complex and its individual components. 2. The comprehensive examination of the hydroxylase substrate specificities utilizing the purified native complex. Substances to be tested will include normal, branched and substituted long-chain alkyl derivatives and steroids. The kinetics of hydroxylation, positional specificity of hydroxylation and the binding affinity will be precisely determined for selected substrates. 3. The identification of the structural and chemical characteristics of the substrate binding sites of the enzyme using primarily affinity-labeling techniques. 4. The characterization of related enzyme systems that act either on the substrates or products of the P-450 monooxygenase system.